Live and Dead Cell Assay (Abcam)

Protocol tips

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	Add 1X Live and Dead Dye to cells. Incubate 10 min at room temperature

Protocol tips
Add 1X Live and Dead Dye to cells. Incubate 10 min at room temperature

Publication protocol

The anti-oomycete assays were adjusted to the characteristics and life-cycles of the oomycete species. Zoospores of S. parasitica and S. diclina were adjusted to 10000 cells/ml and exposed to increasing concentrations (5, 10, 30, 100 µg/ml final concentration) of proteins. Because of the rapid encystment of zoospores into cysts, assessment of mortality was only performed at 30 min (prior to encystment of live zoospores). Zoospores of Phytophthora parasitica were adjusted to 500000 cells/ml and exposed to identical concentrations of proteins (5, 10, 30, 100 ug/ml final concentration). Microscopic observations of the number of live and dead zoospores were performed after 10, 30, 60, 120, 240 min of treatment. Zoospore-treated suspensions were stained immediately after incubation with the Live/Dead Cell Assay kit (Abcam) as described in the manufacturer's protocol. All conditions were tested in triplicate and assays were performed 3 times independently.

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Manufacturer protocol

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